Plant-based alternative to yoghurt

Abstract

The present invention relates to a composition comprising Pediococcus pentosaceus strains. The invention also relates to a starter culture comprising Pediococcus pentosaceus strains and use of Pediococcus pentosaceus strains as a starter culture for fermentation. The present invention also relates to a fermented plant protein, a composition comprising a plant protein and Pediococcus and method of producing the fermented plant protein or composition. The invention also relates to a food product comprising the fermented plant protein or composition.

Description

[0001] PLANT-BASED ALTERNATIVE TO YOGHURT

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] [1] This application claims the benefit of European Application No. 24167810.1, filed March 28, 2024, which is incorporated by reference herein in its entirety.

[0004] FIELD OF THE INVENTION

[0005] [2] The present invention relates to a composition comprising Pediococcus pentosaceus strains, a starter culture comprising Pediococcus pentosaceus strains, use of Pediococcus pentosaceus strains as a starter culture for fermentation, a fermented plant protein, composition and method of producing the same, and a food product comprising the fermented plant protein or composition.

[0006] BACKGROUND OF THE INVENTION

[0007] [3] Yoghurt is a fermented dairy product, produced by bacterial fermentation of animal milk, such as cow’s milk. Bacterial cultures, known as starter cultures, are used to ferment lactose within animal milk. Traditional starter cultures for fermenting lactose utilise Streptococcus thermophilus and Lactobacillus Bulgaricus. These bacteria produce lactic acid, which decreases the pH of the milk causing the lactose to coagulate to form yoghurt. The metabolites produced by the starter cultures affect the texture and flavour of the yoghurt.

[0008] [4] Due to allergies, environmental and ethical concerns, there is an increasing desire for plant-based (vegan) or dairy-free alternatives to yoghurt, which do not contain animal milk or animal protein. However, developing plant-based (vegan) or dairy-free alternatives to yoghurt with suitable texture and sensory properties that mimic traditional yoghurt has proved challenging. For example, the traditional yoghurt starter cultures using Streptococcus thermophilus and Lactobacillus Bulgaricus produce volatile organic compounds which have an undesirable impact on the sensory or taste of plant-based (vegan) or dairy -free alternatives to yoghurt.

[0009] [5] Some plant-based (vegan) or dairy free-alternative to yoghurt utilise legume proteins and non-animal based sugars, instead of milk proteins and lactose. However, consumer acceptance of legume proteins is hindered by unpleasant off-notes or off-flavour (e.g. pea-beany flavours) in the fermented composition caused by the legume proteins.

[0010] [6] Hence, there is a need for an improved tasting plant-based (vegan) or dairy-free alternative to yoghurt. SUMMARY OF THE INVENTION

[0011] [7] The present invention relates to a composition comprising Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545.

[0012] [8] The present invention also relates to a composition comprising Pediococcus pentosaceus C25R2 strain obtained or derived BCCM Accession No. LMG P-33546.

[0013] [9] The present invention relates to a starter culture for fermentation comprising Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545.

[0014]

[0010] The present invention also relates to a starter culture for fermentation comprising Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P- 33546.

[0015]

[0011] The present invention also relates to the use of Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545 as a starter culture for fermentation.

[0016]

[0012] The present invention also relates to the use of Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546 as a starter culture for fermentation.

[0017]

[0013] The starter culture may be a starter culture for fermentation of a foodstuff. The foodstuff may be a dairy alternative (i.e. dairy free). The dairy alternative may be a yoghurt alternative.

[0018]

[0014] The present invention also relates to a fermented plant protein, wherein the plant protein is fermented by lactic acid bacteria, wherein the lactic acid bacteria comprises Pediococcus.

[0019]

[0015] The present invention also relates to a composition comprising: plant protein; and lactic acid bacteria, wherein the lactic acid bacteria comprises Pediococcus.

[0020]

[0016] The plant protein may be derived from legumes. The plant protein may be a pea protein. The pea protein may be a pea protein isolate. The pea protein isolate may be a hydrolysed pea protein. The pea protein may be a pea protein concentrate.

[0021]

[0017] The Pediococcus may be a Pediococcus pentosaceus. The Pediococcus may be the Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545. The Pediococcus may be the Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546.

[0022]

[0018] The lactic acid bacteria may further comprises lactic acid bacteria selected from the group consisting of: Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus , Lactiplantibacillus plantarum and any combination thereof.

[0023]

[0019] The composition of the invention may further comprise a sugar or sugars, a fat or fats, a liquid or liquids, or a combination thereof. The liquid may be water, syrup, juice or a plant-based or non-dairy alternative to milk. The liquid may be water. The sugar may be selected from the group consisting of sucrose, glucose, dextrose, fructose, maltose, lactose and / or galactose and any combination thereof. The fat may be selected from the group consisting of vegetable fat or vegetable oil. The vegetable oil may be coconut oil, sunflower oil, rapeseed oil, palm oil, shea oil, cocoa oil and any combination thereof.

[0024]

[0020] The composition may further comprise one or more emulsifiers, flavourings, thickeners, colourings, additives, probiotics or a combination thereof.

[0025]

[0021] The composition may be a fermented composition.

[0026]

[0022] The plant protein of the composition may be a fermented plant protein.

[0027]

[0023] The composition may have one or more of the following characteristics: a. 0.5-12 grams of protein per 100 grams of composition; b. 1-15 grams of carbohydrates per 100 grams of composition; c. 0-10 grams of fat per 100 grams of composition; and / or d. 0-150 mg of calcium per 100 grams of composition.

[0028]

[0024] The composition may be a foodstuff. The foodstuff may be a yoghurt alternative.

[0029]

[0025] The invention also relates to a process for preparing a fermented plant protein or a composition, the process comprising: a) Providing an aqueous composition comprising a plant protein; b) Providing lactic acid bacteria, wherein the lactic acid bacteria comprises Pediococcus.

[0030]

[0026] The plant protein may be derived from legumes. The plant protein may be a pea protein. The pea protein may be a pea protein isolate. The pea protein isolate may be a hydrolysed pea protein. The pea protein may be a pea protein concentrate.

[0031]

[0027] The Pediococcus may be Pediococcus pentosaceus. The Pediococcus may be the Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545. The Pediococcus may be the Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546.

[0032]

[0028] The process may be a process for preparing a fermented plant protein and step (b) may comprise fermenting the composition with the lactic acid bacteria to provide a fermented plant protein.

[0033]

[0029] Step (b) may be carried out at 25-45°C. Step (b) may be carried out at 28°C.

[0030] The lactic acid bacteria may further comprise lactic acid bacteria selected from the group consisting of: Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactiplantibacillus plantarum and any combination thereof. The Pediococcus pentosaceus may be combined with Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactiplantibacillus plantarum and any combination thereof in a 50:50 ratio. Step (b) may be carried out at 25-45°C. Step (b) may be carried out at 28-37°C.

[0034]

[0031] The process may be a process for preparing a composition. Step (b) may be carried out at 25-45°C. Step (b) may be carried out at 28°C. The lactic acid bacteria in step (b) may ferment the aqueous composition comprising a plant protein.

[0035]

[0032] The lactic acid bacteria may be selected from the group consisting of: Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactiplantibacillus plantarum and any combination thereof, the Pediococcus pentosaceus may be combined with Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactiplantibacillus plantarum and any combination thereof in a 50:50 ratio. Step (b) may be carried out at 25-45°C. Step (b) may be carried out at 28-37°C.

[0036]

[0033] The plant protein may be hydrated. The composition may be subjected to mixing after step (a) but before step (b), such as high shear mixing. The composition may be homogenized after step (a) but before step (b). The composition may be subjected to a heat treatment after step (a) but before step (b) to pasteurise the composition. The heat treatment may be 2-10 minutes at atemperature of 80-100°C. The composition may be cooled before step (b). The composition may be cooled to the fermentation temperature of the lactic acid bacteria. The composition may be cooled to 25-45°C. The process may further comprises (c) smoothening the composition.

[0034] Step (a) of the process may further comprise adding one or more additional ingredients to the aqueous composition comprising a plant protein. The one or more additional ingredients may include a sugar or sugars, a fat or fats, a liquid or liquids, emulsifiers, flavourings, thickeners, colourings, additives, probiotics or a combination thereof. The liquid may be water, syrup, juice or a plant-based or non-dairy alternative to milk. The liquid may be water. The sugar may be selected from the group consisting of sucrose, glucose, dextrose, fructose, maltose, lactose and / or galactose and any combination thereof. The fat may be selected from the group consisting of vegetable fat or vegetable oil. The vegetable oil may be coconut oil, sunflower oil, rapeseed oil, palm oil, shea oil, cocoa oil and any combination thereof.

[0037]

[0035] The invention also relates to a food product comprising the fermented plant protein as described herein or the composition as described herein. The product may be a dairy alternative. The dairy alternative may be a yoghurt alternative.

[0038]

[0036] Representative features of the present invention are set out in the following clauses, which stand alone or may be combined, in any combination, with one or more features disclosed in the text and / or figures of the specification.

[0039]

[0037] The present invention is as set out in the following clauses:

[0040] 1. A composition comprising Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545.

[0041] 2. A composition comprising Pediococcus pentosaceus C25R2 strain obtained or derived BCCM Accession No. LMG P-33546.

[0042] 3. A starter culture for fermentation comprising Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545 or Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546.

[0043] 4. The starter culture of clause 3, wherein the starter culture is a starter culture for fermentation of a foodstuff.

[0044] 5. The starter culture of clause 4, wherein the foodstuff is a dairy alternative.

[0045] 6. The starter culture of clause 5, wherein the dairy alternative is a yoghurt alternative.

[0046] 7. Use of Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545 or Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546 as a starter culture for fermentation.

[0047] 8. The use of clause 7, wherein the starter culture is a starter culture for fermentation of a foodstuff.

[0048] 9. The use of clause 8, wherein the foodstuff is a dairy alternative.

[0049] 10. The use of clause 9, wherein the dairy alternative is a yoghurt alternative. A fermented plant protein, wherein the plant protein is fermented by lactic acid bacteria, wherein the lactic acid bacteria comprises Pediococcus. The fermented plant protein of clause 11, wherein the plant protein is derived from legumes. The fermented plant protein of clause 12, wherein the plant protein is a pea protein. The fermented plant protein of clause 13, wherein the pea protein is a pea protein isolate. The fermented plant protein of clause 14, wherein the pea protein isolate is a hydrolysed pea protein. The fermented plant protein of clause 13, wherein the pea protein is a pea protein concentrate. The fermented plant protein of clauses 11-16, wherein the Pediococcus is Pediococcus pentosaceus. The fermented plant protein of clause 17, wherein the Pediococcus pentosaceus is Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545. The fermented plant protein of clause 17, wherein the Pediococcus pentosaceus is Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546. The fermented plant protein of any one of clauses 11-19, wherein the lactic acid bacteria further comprises lactic acid bacteria selected from the group consisting of: Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactiplantibacillus plantarum and any combination thereof. A process for preparing a fermented plant protein, the process comprising: a) Providing an aqueous composition comprising a plant protein; b) Fermenting the composition with lactic acid bacteria to provide a fermented plant protein, wherein the lactic acid bacteria comprises Pediococcus. The process of clause 21, wherein the plant protein is derived from legumes. The process of clause 22, wherein the plant protein is a pea protein. The process of clause 23, wherein the plant protein is a pea protein isolate. The process of clause 24, wherein the pea protein isolate is a hydrolysed pea protein. The process of clause 23, wherein the pea protein is a pea protein concentrate. The process of clauses 21-26, wherein the Pediococcus is Pediococcus pentosaceus. The process of clause 27, wherein the Pediococcus pentosaceus is Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P- 33545. The process of clause 27, wherein the Pediococcus pentosaceus is Pediococcus pentosaceus C25R2 obtained or derived from BCCM Accession No. LMG P-33546. The process of clauses 21-29, wherein step (b) is carried out at 25-45°C. The process of clause 30, wherein step (b) is carried out at 28°C. The process of clauses 21-31, wherein the lactic acid bacteria of step (b) further comprises lactic acid bacteria selected from the group consisting of: Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactiplantibacillus plantarum and any combination thereof. The process of clause 32, wherein Pediococcus pentosaceus and Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactiplantibacillus plantarum and any combination thereof are provided in a 50:50 ratio. The process of clauses 32 or 33, wherein step (b) is carried out at 25-45°C. The process of clause 34, wherein step (b) is carried out at 28-37°C. A composition comprising: plant protein; and lactic acid bacteria, wherein the lactic acid bacteria comprises Pediococcus. The composition of clause 36, wherein the plant protein is derived from legumes. The composition of clause 37, wherein the plant protein is a pea protein. The composition of clause 38, wherein the pea protein is pea protein isolate. The composition of clause 39, wherein the pea protein isolate is a hydrolysed pea protein. The composition of clause 38, wherein the pea protein is a pea protein concentrate. The composition of clauses 36-41, wherein the Pediococcus is Pediococcus pentosaceus. The composition of clause 42, wherein the Pediococcus pentosaceus is Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P- 33545. The composition of clause 42, wherein the Pediococcus pentosaceus is Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546. The composition of any one of clauses 36-44, wherein the lactic acid bacteria further comprises lactic acid bacteria selected from the group consisting of: Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactiplantibacillus plantarum and any combination thereof. The composition of any one of clauses 36-45, wherein the composition further comprises a sugar or sugars, a fat or fats, a liquid or liquids, or a combination thereof. The composition of clause 46, wherein the liquid is water, syrup, juice or a plant-based or non-dairy alternative to milk. The composition of clause 47, wherein the liquid is water. The composition of clause 46, wherein the sugar is selected from the group consisting of sucrose, glucose, dextrose, fructose, maltose, lactose and / or galactose and any combination thereof. The composition of clause 49, wherein the fat is selected from the group consisting of vegetable fat or vegetable oil. The composition of clause 50, wherein the vegetable oil is coconut oil, sunflower oil, rapeseed oil, palm oil, shea oil, cocoa oil and any combination thereof. The composition of any one of clauses 36-51, further comprising one or more emulsifiers, flavourings, thickeners, colourings, additives, probiotics or a combination thereof. The composition of any one of clauses 36-52, wherein the composition is a fermented composition. The composition of any one of clauses 36-53, wherein the plant protein is a fermented plant protein. The composition of any one of clauses 36-54, wherein the composition has one or more of the following characteristics:

[0050] (i) 0.5-12 grams of protein per 100 grams of composition;

[0051] (ii) 1-15 grams of carbohydrates per 100 grams of composition;

[0052] (iii) 0-10 grams of fat per 100 grams of composition; and / or

[0053] (iv) 0-150 mg of calcium per 100 grams of composition. The composition of any one of clauses 36-55, wherein the composition is a foodstuff. The composition of clause 56, wherein the foodstuff is a yoghurt alternative. A process for preparing a composition, the process comprising: a) Providing an aqueous composition comprising a plant protein; b) Providing lactic acid bacteria, wherein the lactic acid bacteria comprises Pediococcus. The process of clause 58, wherein the plant protein is derived from legumes. The process of clause 59, wherein the plant protein is a pea protein. The process of clause 60, wherein the pea protein is a pea protein isolate. The process of clause 61, wherein the pea protein isolate is a hydrolysed pea protein. The process of clause 60, wherein the pea protein is a pea protein concentrate. The process of clauses 58-63, wherein the Pediococcus is Pediococcus pentosaceus. The process of clause 64, wherein the Pediococcus pentosaceus is Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P- 33545. The process of clause 64, wherein the Pediococcus pentosaceus is Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546. The process of clauses 58-66 wherein step (b) is carried out at 25-45°C. The process of clause 67, wherein step (b) is carried out at 28°C. The process of clauses 58-68, wherein the lactic acid bacteria in step (b) ferments the aqueous composition comprising a plant protein. The process of clauses 58-69, further comprising providing lactic acid bacteria selected from the group consisting of: Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactiplantibacillus plantarum and any combination thereof. The process of clause 70, wherein Pediococcus pentosaceus and Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactiplantibacillus plantarum and any combination thereof are provided in a 50:50 ratio. The process of clauses 70 or 71, wherein step (b) is carried out at 25-45°C. The process of clause 72, wherein step (b) is carried out at 28-37°C. The process of clauses 58-73, wherein the plant protein is hydrated. The process of any one of clauses 58-74, wherein the composition is subjected to mixing after step (a) but before step (b). The process of any one of clauses 58-75, wherein the composition is homogenized after step (a) but before step (b). The process of any one of clauses 58-76, wherein the composition is subjected to a heat treatment after step (a) but before step (b) to pasteurise the composition. The process of clause 77, wherein the heat treatment is 2-10 minutes at a temperature of 80-100°C. The process of any one of clauses 58-78, wherein the composition is cooled before step (b). The process of clause 79, wherein the composition is cooled to the fermentation temperature of the lactic acid bacteria. The process of clause 79, wherein the composition is cooled to 25-45°C. The process of any one of clauses 58-82, wherein the process further comprises

[0054] (c) smoothening the composition. The process of any one of clauses 58-83, wherein step (a) further comprises adding one or more additional ingredients to the aqueous composition comprising a plant protein. The process of clause 83, wherein the one or more additional ingredients include a sugar or sugars, a fat or fats, a liquid or liquids, emulsifiers, flavourings, thickeners, colourings, additives, probiotics or a combination thereof. The process of clause 84, wherein the liquid is water, syrup, juice or a plant-based or nondairy alternative to milk. The process of clause 85, wherein the liquid is water. The process of clause 84, wherein the sugar is selected from the group consisting of sucrose, glucose, dextrose, fructose, maltose, lactose and / or galactose and any combination thereof. The process of clause 84, wherein the fat is selected from the group consisting of vegetable fat or vegetable oil. The process of clause 84, wherein the vegetable oil is coconut oil, sunflower oil, rapeseed oil, palm oil, shea oil, cocoa oil and any combination thereof. A food product comprising the fermented plant protein of any one of clauses 11-20 or the composition of any one of clauses 36-57. The food product of clause 90, wherein the product is a dairy alternative. The food product of clause 91, wherein the dairy alternative is a yoghurt alternative. DETAILED DESCRIPTION

[0055]

[0038] Embodiments of the invention are described below with reference to the accompanying drawings. The accompanying drawings illustrate various embodiments of systems, methods, and embodiments of various other aspects of the disclosure. Any person with ordinary skills in the art will appreciate that the illustrated element boundaries (e.g., boxes, groups of boxes, or other shapes) in the figures represent one example of the boundaries. It may be that in some examples one element may be designed as multiple elements or that multiple elements may be designed as one element. In some examples, an element shown as an internal component of one element may be implemented as an external component in another and vice versa. Furthermore, elements may not be drawn to scale. Non-limiting and non-exhaustive descriptions are described with reference to the following drawings. The components in the figures are not necessarily to scale, emphasis instead being placed upon illustrating principles.

[0056]

[0039] Figure 1 is a sensory analysis of pea protein fermented with Pediococcus pentosaceus C25R2 strain (Fermentate 1) and Pediococcus pentosaceus C22B17 strain (Fementate 2) according to examples of the invention.

[0057]

[0040] Figure l is a non-volatile principle component analysis (PCA) graph of water extract from Fermentate 1.

[0058]

[0041] Figure 3 is a volcano plot of water extract from LAB fermentation samples from Fermentate 1. Oh fermented versus 24 hours fermented. P-value is 0.01 and Log2 fold change is 1.

[0059]

[0042] Figure 4 is a non-volatile PCA graph of organic / water extract from LAB fermentation samples from Fermentate 1 .

[0060]

[0043] Figure 5 is a volcano plot of water / organic extract from LAB fermentation samples from Fermentate 1. Oh fermented versus 24 hours fermented. P-value is 0.05 and Log2 fold change is 0.5.

[0061]

[0044] Figure 6 is a line graph showing the sensory results concerning smell of a plant-based alternative to yoghurt according to the present invention fermented with Pediococcus pentosaceus C22B17compared to a plant-based alternative to yoghurt which has been fermented with a commercial starter culture (VegaVibe™) and a plant-based alternative to yoghurt which has been fermented with a mixture of Pediococcus pentosaceus C22B17 and a commercial starter culture (VegaVibe™).

[0062]

[0045] Figure 7 is a line graph showing the sensory results concerning taste / flavour of a plantbased alternative to yoghurt according to the present invention fermented with Pediococcus pentosaceus C22B17compared to a plant-based alternative to yoghurt which has been fermented with a commercial starter culture (VegaVibe™) and a plant-based alternative to yoghurt which has been fermented with a mixture of Pediococcus pentosaceus C22B17 and a commercial starter culture (VegaVibe™).

[0063]

[0046] Figure 8 is a line graph showing the sensory results concerning taste / flavour of a plantbased alternative to yoghurt according to the present invention fermented with Pediococcus pentosaceus C22B17compared to a plant-based alternative to yoghurt which has been fermented with a commercial starter culture (VegaVibe™) and a plant-based alternative to yoghurt which has been fermented with a mixture of Pediococcus pentosaceus C22B17 and a commercial starter culture (VegaVibe™).

[0064]

[0047] Figure 9 is a line graph showing the sensory results concerning texture of a plant-based alternative to yoghurt according to the present invention fermented with Pediococcus pentosaceus C22B17 compared to a plant-based alternative to yoghurt which has been fermented with a commercial starter culture (VegaVibe™) and a plant-based alternative to yoghurt which has been fermented with a mixture of Pediococcus pentosaceus C22B17 and a commercial starter culture (VegaVibe™).

[0065]

[0048] Figure 10 is a line graph showing the sensory results concerning afterfeel of a plant-based alternative to yoghurt according to the present invention fermented with Pediococcus pentosaceus C22B17 compared to a plant-based alternative to yoghurt which has been fermented with a commercial starter culture (VegaVibe™) and a plant-based alternative to yoghurt which has been fermented with a mixture of Pediococcus pentosaceus C22B17 and a commercial starter culture (VegaVibe™).

[0066]

[0049] Embodiments of the present disclosure will be described more fully hereinafter with reference to the accompanying drawings in which like numerals represent like elements throughout the several figures, and in which example embodiments are shown. Embodiments of the claims may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

[0067]

[0050] The words "comprising," "having," "containing," and "including," and other forms thereof, are intended to be equivalent in meaning and be open ended in that an item or items following any one of these words is not meant to be an exhaustive listing of such item or items or meant to be limited to only the listed item or items. It must also be noted that as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Although any systems and methods similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present disclosure, the preferred systems and methods are now described.

[0068]

[0051] Some of the terms used to describe the present invention are set out below:

[0052] “ Starter culture” refers to cells from a single microorganism or more than one microorganism which are added to a substrate and ferment the substrate. The starter culture may include cells from one or more species or strains of lactic acid bacteria (LAB), such as, for example, from the genus Lactococcus, Lactobacillus, Leuconostoc and Pediococcus and any combination thereof. Preferably, the starter culture includes cells from Pediococcus pentosaceus. The added starter culture may contain 4-6 log cells / gram or cells / millilitre.

[0069]

[0053] “Lactic acid bacteria” refers to bacteria which produce lactic acid when metabolising carbohydrates, and includes bacteria from the genus Lactococcus, Lactobacillus, Leuconostoc and Pediococcus and any combination thereof.

[0070]

[0054] “Fermented composition” refers to a composition where the carbohydrates of such composition have been converted to alcohol or organic acids using microorganisms, such as bacteria, under anaerobic conditions.

[0071]

[0055] “Yoghurt” refers to a food or drink product formed by the conversion of lactose in animal milk, such as cow’s milk to lactic acid by a starter culture containing lactic acid bacteria, commonly Streptococcus thermophilus and Lactobacillus delbrueckii subsp. Bulgaricus. The term “yoghurt” encompasses any composition made with animal milk, including traditional yoghurt, flavoured yoghurt, kefir yoghurt, Greek yoghurt, Labneh, and Skyr (Icelandic) yoghurt. The term “Yoghurt” encompasses fermented compositions.

[0072]

[0056] “Plant-based alternative” or “vegan” refers to an alternative, mimic, substitute or analog for food or drink products traditionally utilising animal products, which does not contain any ingredients obtained or derived from animals, such as animal milk or animal protein, such as cow’s milk and are suitable for a vegan diet. This includes but is not limited to a plant-based alternative, mimic, substitute or analog of yoghurt or vegan yoghurt, which encompasses edible products that are designed to mimic traditional yoghurts in terms of properties such as, but not limited to appearance, texture and / or taste; but wherein constituents including milk fat and / or milk proteins have been replaced by ingredients that are not originating from animals. The terms alternative, mimic, substitute or analog are used interchangeably herein and mean a plant-based or vegan replacement to yoghurt that does not comprise any ingredient that is originating from animals. The alternative is a plant-based or vegan alternative of any form of yoghurt, including an alternative to traditional yoghurt, flavoured yoghurt, kefir yoghurt, Labneh, Greek yoghurt and Skyr (Icelandic) yoghurt.

[0073]

[0057] “Dairy alternative” refers to a dairy free alternative for products traditionally based on animal milk, such as but not limited to non-dairy alternatives for yoghurt. The terms alternative, mimic, substitute or analog are used interchangeably herein and mean a dairy free replacement to yoghurt (traditionally made with animal milk). The alternative is a dairy-free alternative of any form of yoghurt, including alternative to kefir yoghurt, Greek yoghurt and Skyr (Icelandic) yoghurt.

[0074]

[0058] “Foodstuff’ refers to foods intended for eating and beverages intended for drinking, including, but not limited to, alternatives, analogues and substitutes of dairy products, including yoghurts, yoghurt beverages and yoghurt containing food products.

[0075] Lactic acid bacteria

[0076]

[0059] The invention utilises lactic acid bacteria, which metabolise carbohydrates to produce lactic acid.

[0077]

[0060] The present invention utilises Pediococcus, a genus of gram-positive LAB within the family of Lactobacillaceae. The Pediococcus may be used as a starter culture for fermentation. The Pediococcus may be used as a starter culture to produce yoghurt, particularly a plant-based alternative, vegan or dairy-free alternative to yoghurt. The Pediococcus may be used to ferment a plant protein, such as a legume protein, such as a pea protein.

[0078]

[0061] The Pediococcus may be Pediococcus pentosaceus. The Pediococcus pentosaceus may bet the Pediococcus pentosaceus C22B17 strain. The Pediococcus pentosaceus may be the Pediococcus pentosaceus C25R2 strain.

[0079]

[0062] “Pediococcus pentosaceus C22B17 strain" refers to the microorganism strain deposited at Belgian Coordinated Collections of Micro-organisms (BCCM), Laboratorium voor Microbiol ogie

[0080] - Bacterienverzameling (LMG), Universiteit Gent, K.L. Ledeganckstraat 35, B-9000 Gent on February 15th2024 under deposit number LMG P-33545 by Cargill Incorporated, 15407 McGinty Road West, MS163, Wayzata, MN 55391, USA. This strain was isolated from a buckwheat sourdough. In particular, this strain was isolated from a sourdough elaborated with organic buckwheat flour and dough yield (DY) 200, which was incubated at 22°C for 24h.

[0081]

[0063] “Pediococcus pentosaceus C25R2 strain" refers to the microorganism strain deposited at Belgian Coordinated Collections of Micro-organisms (BCCM), Laboratorium voor Microbiol ogie

[0082] - Bacterienverzameling (LMG), Universiteit Gent, K.L. Ledeganckstraat 35, B-9000 Gent on February 15th2024 under deposit number LMG P-33546 by Cargill Incorporated, 15407 McGinty Road West, MS163, Wayzata, MN 55391, USA. This strain was isolated from a rye sourdough. In particular, this strain was isolated from a sourdough elaborated with organic rye flour and dough yield (DY) 200, which was incubated at 25°C for 24h and was back-slopped once following the same conditions.

[0083]

[0064] The present inventors have discovered that Pediococcus pentosaceus, in particular, Pediococcus pentosaceus C22B 17 strain and Pediococcus pentosaceus C25R2 strain can be used to ferment legume proteins, such as pea proteins, and provides improved tasting fermentation product with reduced off-flavours, such as reduced beany or pea flavours. Without wishing to be bound by theory, the present inventors have discovered that through fermentation Pediococcus pentosaceus can reduce chemical compounds such as aldehyde, ketones that are associated with green, beany notes, and increase alcohols, that are more fruity, mild and / or pleasant.

[0084]

[0065] The present invention also provides a composition comprising Pediococcus pentosaceus C22B17 strain. The present invention also provides a composition comprising Pediococcus pentosaceus C25R2 strain. The composition may comprise the Pediococcus pentosaceus C22B17 strain and the Pediococcus pentosaceus C25R2 strain.

[0085]

[0066] The Pediococcus pentosaceus C22B17 strain and / or the Pediococcus pentosaceus C25R2 strain may be used as a starter culture for fermentation of a protein, such as a plant protein.

[0086]

[0067] The present invention encompasses a starter culture for fermentation of a foodstuff comprising Pediococcus pentosaceus C22B17 strain and / or Pediococcus pentosaceus C25R2 strain.

[0087]

[0068] The lactic acid bacteria may further comprise lactic acid bacteria selected from the group consisting of Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Pediococcus acidilactici, Lactiplantibacillus plantarum and any combination thereof. The lactic acid bacteria may comprise Pediococcus pentosaceus C22B17 strain in combination with Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Pediococcus acidilactici or Lactiplantibacillus plantarum, or any combination thereof The lactic acid bacteria may comprise Pediococcus pentosaceus C25R2 strain in combination with Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Pediococcus acidilactici or Lactiplantibacillus plantarum, or any combination thereof The lactic acid bacteria may comprise Pediococcus pentosaceus C22B17 strain and Pediococcus pentosaceus C25R2 strain in combination with Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Pediococcus acidilactici, or Lactiplantibacillus plantarum, or any combination thereof. The lactic acid bacteria comprises Pediococcus pentosaceus C22B17 strain and Lactiplantibacillus plantarum.

[0088] Plant protein

[0089]

[0069] The invention includes a fermented plant protein, wherein the plant protein is fermented by LAB, wherein the LAB comprises Pediococcus. The plant protein may be fermented by Pediococcus pentosaceus. The plant protein may be fermented by the Pediococcus pentosaceus C22B17 strain. The plant protein may be fermented by the Pediococcus pentosaceus C25R2 strain. The plant protein may be fermented by the Pediococcus pentosaceus C22B17 strain and the Pediococcus pentosaceus C25R2 strain. The plant protein may be fermented by the Pediococcus pentosaceus C22B17 strain and / or the Pediococcus pentosaceus C25R2 strain and at least one other LAB species.

[0090]

[0070] The present inventors have discovered that Pediococcus, such as Pediococcus pentosaceus, and in particular, the Pediococcus pentosaceus C22B17 strain and the Pediococcus pentosaceus C25R2 strain can be used, e.g. as a starter culture, to ferment plant proteins, such as pea proteins, and the undesirable plant protein (e.g. pea) flavours can be eliminated or covered resulting in a more neutral tasting product.

[0091]

[0071] The plant protein may be derived from a legume. “Legume” refers to pea, soy, fava, chickpea, lentil and lupin.

[0092]

[0072] The plant protein may be a pea protein, i.e. a protein from Pisum sativum. “Pea protein” refers to pea protein isolate, pea protein concentrate, whole pea powder, pea flour, extruded micronized pea powder, hydrolysed pea protein or the like.

[0093]

[0073] The pea protein may be a pea protein isolate. “Pea protein isolates” have a total protein content of 80 to 95 wt% on a dry weight basis.

[0094]

[0074] The pea protein may be a pea protein concentrate. “Pea protein concentrates” have a total protein content of 50 to 75 wt% on a dry weight basis.

[0095]

[0075] The pea protein may be hydrolysed, i.e. is a pea protein hydrolysate. “Pea protein hydrolysates” are preparations obtained by enzymatic and / or chemical hydrolysis of pea protein. Protein hydrolysates consist of a mixture of peptides of different sizes and free amino acids. The pea protein may not be hydrolysed.

[0096]

[0076] The pea protein may be a hydrolysed pea protein isolate. The pea protein may be a pea protein isolate which is not hydrolysed.

[0077] The pea protein may be RadiPure E®, which is a hydrolysed pea protein isolate comprising a protein content of at least 80% on a dry weight basis, RadiPure S®, which is a pea protein isolate comprising a protein content of at least 80% on a dry weight basis, Puris 870H®, which is a hydrolysed pea protein isolate comprising a protein content of at least 80% on a dry weight basis, Puris 870® which is a pea protein isolate comprising a protein content of at least 80% on a dry weight basis or Puris 2.0 ® which is a pea protein isolate comprising a protein content of at least 80% on a dry weight basis.

[0097] Alternative to yoghurt

[0098]

[0078] Compositions according to the invention have various desirable properties, which make these compositions useful, for example, as a plant-based, vegan or dairy free alternative to yoghurt.

[0099]

[0079] The invention provides a composition comprising plant protein and lactic acid bacteria, wherein the lactic acid bacteria comprises Pediococcus. such as Pediococcus pentosaceus. The lactic acid bacteria may ferment the plant protein to produce a fermented composition. The composition may comprise 0.01-0.10 wt %, preferably 0.03 wt % of Pediococcus pentosaceus strains.

[0100]

[0080] The composition may comprise 5-15 wt %, preferably 8 wt % of plant protein, preferably pea protein.

[0101]

[0081] The composition may further comprise a sugar or sugars, a fat or fats, a liquid or liquids, or a combination thereof.

[0102]

[0082] The liquid may be selected from the group consisting of water, syrup such as oat syrup, juice or a plant-based or non-dairy alternative to milk, such as oat milk, soya milk, coconut milk, almond milk or hazelnut milk. The liquid may be water. The liquid in the composition does not include liquid fat or liquid oil. The composition may comprise 70-90 wt %, preferably 80-86 wt % of liquid. The skilled person would appreciate that the amount of liquid, such as water, present in the composition is quantum satis depending on the amount of other ingredients present in the composition.

[0103]

[0083] The sugar may be selected from the group consisting of sucrose, glucose, dextrose, fructose, maltose, lactose and / or galactose and any combination thereof. The composition may comprise 0-10 wt %, preferably 4 wt % of sugar. The objective of adding sugar to the composition is to promote the fermentation and / or to tone down the acidity of the composition. The level of incorporation of sugar may dependent upon consumer preference, type of fruit used, and the like.

[0104]

[0084] The fat may be selected from vegetable fat or vegetable oil. The vegetable oil may be coconut oil, sunflower oil, rapeseed oil, palm oil, shea oil, cocoa oil and any combination thereof. The composition may comprise 0-20 wt%, preferably 0-10 wt %, even more preferably 0-5 wt % of fat.

[0105]

[0085] The composition may further comprise other ingredients that are commonly present in yoghurt, such as, but not limited to one or more emulsifiers, flavourings, thickeners, texturisers, colourings, preservatives, additives, fruits, fruit preparations such as fruit prep, nuts, cereals and the like or a combination thereof. The composition may comprise 0-10 wt % of emulsifiers, flavourings, thickeners, colourings, additives and / or probiotics.

[0106]

[0086] Examples of texturizers are carrageenans, seaweed powder, carboxymethyl celluloses, agar agar, guar gum, xanthan gum, locust bean gum, gum arabic, basil seed gum, pectin, sodium alginate, gellan, starches in native or modified forms from wheat, maize, rice, potato, com, tapioca, maltodextrin, fiber or a combination of two or more thereof. The texturizers may be present in an amount of from 0 to 6 wt.%, on total weight of the composition. Alternatively, no texturizer may be present.

[0107]

[0087] The composition may further comprise functional ingredients such as, but not limited to, vitamins and / or inerals, bioactive compounds, dietary fiber, probiotics, prebiotics, or a combination thereof.

[0108]

[0088] The composition may be a fermented composition, preferably wherein the composition is fermented by Pediococcus.

[0109]

[0089] The composition may have one or more of the following characteristics:

[0110] (v) 0.5-12 grams of protein per 100 grams of composition.

[0111] (vi) 1-15 grams of carbohydrates per 100 grams of composition

[0112] (vii) 0-10 grams of fat per 100 grams of composition; and / or

[0113] (viii) 0-95 mg of calcium per 100 grams of composition

[0114]

[0090] The protein may comprise a plant protein, such as a pea protein as described herein.

[0115]

[0091] The composition may have a pH of more than 4.6. The composition may have a pH of 4.6. The composition may have a pH of 4.7. The composition may have a pH of 4.7.

[0116] Processes

[0117]

[0092] The invention also provides a process for preparing a composition as described herein, such as a fermented composition, or a process for preparing a fermented plant protein as described herein.

[0118]

[0093] The process may comprise a) providing an aqueous composition comprising a plant protein and b) providing lactic acid bacteria, wherein the lactic acid bacteria comprises Pediococcus. The lactic acid bacteria may comprise Pediococcus pentosaceus, in particular the Pediococcus pentosaceus C22B17 strain and / or the Pediococcus pentosaceus C25R2 strain as described herein. The lactic acid bacteria may ferment the plant protein to provide a fermented plant protein or a fermented composition as described herein. The plant protein is a plant protein as described herein.

[0119]

[0094] The aqueous composition may comprise water, syrup such as oat syrup, juice or a plantbased or non-dairy alternative to milk, such as oat milk, soya milk, coconut milk, almond milk or hazelnut milk. The aqueous composition may not comprise liquid fat or liquid oil. The composition may comprise 70-90 wt %, preferably 80-86 wt % of liquid. The skilled person would appreciate that the amount of liquid, such as water, present in the composition is quantum satis depending on the amount of other ingredients present in the composition.

[0120]

[0095] The lactic acid bacteria may be provided at their fermentation temperature. For example, the fermentation temperature of Pediococcus pentosaceus is 25-45°C. Therefore, the lactic acid bacteria may be provided at 25-45°C. The lactic acid bacteria may be provided at 25-30°C. The lactic acid bacteria may be provided at 28°C, which is the optimum fermentation temperature of Pediococcus pentosaceus C22B17 strain and Pediococcus pentosaceus C25R2 strain.

[0121]

[0096] The process may further comprise providing additional lactic acid bacteria, such as Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus and any combination thereof as described herein. The additional lactic acid bacteria may be provided at 25-45°C. The additional lactic acid bacteria may be provided at 28-37°C. Ideally, the additional lactic acid bacteria are provided at their optimum fermentation temperature.

[0122]

[0097] The Pediococcus pentosaceus, such as the Pediococcus pentosaceus C22B17 strain and / or the Pediococcus pentosaceus C25R2 strain and the additional lactic acid bacteria may be provided in a 50:50 ratio. The Pediococcus pentosaceus, such as the Pediococcus pentosaceus C22B17 strain or the Pediococcus pentosaceus C25R2 strain and the additional lactic acid bacteria may be provided in a 90: 10 or a 10:90 ratio. The Pediococcus pentosaceus, such as the Pediococcus pentosaceus C22B17 strain or Pediococcus pentosaceus C25R2 strain and the additional lactic acid bacteria may be provided in an 80:20 or a 20:80 ratio. The Pediococcus pentosaceus, such as the Pediococcus pentosaceus C22B17 strain or the Pediococcus pentosaceus C25R2 strain and the additional lactic acid bacteria may be provided in an 70:30 or a 30:70 ratio. The Pediococcus pentosaceus, such as the Pediococcus pentosaceus C22B1 / strain or the Pediococcus pentosaceus C25R2 strain and the additional lactic acid bacteria may be provided in an 60:40 or a 60:40 ratio. The ratio of the different lactic acid bacteria may be adjusted based on the fermentation temperature of the composition, because particular temperatures may favour a particular type of lactic acid bacteria to grow faster compared to the other, hence become the more dominant species in the final PBAY

[0123]

[0098] The plant protein (e.g. pea protein) may be hydrated. Pea protein hydration can improve the protein functionality (e.g. the acid induced gelation in yoghurt) and solubility of the composition. The plant protein (e.g. plant protein) may be hydrated in water at a temperature of 1-85°C for a time period of between 2 minutes to 24 hours. For example, the plant protein (e.g. pea protein) may be hydrated at 4 °C for 24 hours. The plant protein (e.g. pea protein) may be hydrated at 65°C for 15 minutes. The plant protein (e.g. pea protein) may be hydrated at 85°C for 5 minutes. Alternatively, the plant protein may not be hydrated.

[0124]

[0099] The composition may be subjected to mixing after step (a) but before step (b). This provides a homogenous composition without any lumps in the composition. Any form of mixing known in the art to provide a homogenous composition can be used. The mixing may be high shear mixing.

[0125]

[0100] The composition may be homogenised after step (a) but before step (b). Homogenisation makes an emulsion of fat in water. If no fat is present in the composition, then no homogenisation is required. Homogenisation may comprise applying a high pressure, such as a pressure of 100- 250 bar, to the composition. The temperature of the composition during homogenisation depends on the fat or oil being used in the composition. The homogenisation may be carried out above the melting temperature of the fat or oil.

[0126]

[0101] The composition may be subjected to a heat treatment after step (a) but before step (b). This pasteurises or sterilises the composition to eliminate pathogens and extend shelf life. The heat treatment may be 2-10 minutes at a temperature of 80-100°C. The heat treatment may be 5 minutes at 95°C.

[0127]

[0102] The composition may be cooled before step (b). The composition may be cooled to the fermentation temperature of the lactic acid bacteria in the starter culture. The composition may be cooled to 25-45°C.

[0128]

[0103] The composition may remain at the fermentation temperature of the lactic acid bacteria during step (b).

[0129]

[0104] Step (b) may be a fermentation step where the lactic acid bacteria ferments the plant protein in the composition to produce a fermented composition. The composition may be cooled to stop fermentation once the pH of the composition is between 4.6-4.8, or lower than 4.6.

[0130]

[0105] The composition may be smoothened after step (b). This breaks up the set gel formed by fermentation. The composition may be smoothened using low shear. The composition may be pumped through a sieve.

[0131]

[0106] The method may comprise the following steps: (i) Optionally hydrating the plant protein, and providing an aqueous composition comprising a plant protein

[0132] (ii) Optional mixing of the composition

[0133] (iii) Optional homogenization of the composition

[0134] (iv) Optional pasteurisation of the composition

[0135] (v) Providing lactic acid bacteria to ferment the plant protein to form a yoghurt alternative composition

[0136] (vi) Optionally smoothening the yoghurt alternative composition.

[0137] Foodstuff

[0138]

[0107] The starter culture may be a starter culture for fermentation of a foodstuff.

[0139]

[0108] The fermented plant protein may be a fermented plant protein as part of a foodstuff.

[0140]

[0109] The composition may be a foodstuff.

[0141]

[0110] The foodstuff may be a dairy alternative, such as a yoghurt alternative. The yoghurt alternative may be a spoonable or drinkable alternative yoghurt, or a ready-to-drink alternative yoghurt beverage.

[0142] EXAMPLES

[0143]

[0111] The following are non-limiting examples that discuss, with reference to tables and figures, the advantages of the present invention. The examples set forth herein are merely examples among other possible examples.

[0144]

[0112] Example 1: Culture isolation ofPediococcus pentosaceus C22B17 strain and Pediococcus pentosaceus C25R2 strain

[0145]

[0113] The spontaneous sourdough fermentations were samples following preparation of serial dilutions in sterile physiological peptone solution (PPS; 0.85% NaCl and 0.1% bacteriological peptone) and spread-plated on different agar media. The main laboratory medium used was de Man-Rogosa- Sharpe (MRS) using sucrose, glucose or lactose as carbon source.

[0146] Table 1. MRS Composition

[0147]

[0114] The plates were incubated at 22°C for 5 days and single colonies were picked up at the end of the incubation period based on colony morphology and aspect of the cells under the microscope. Pure colonies were picked up, transferred in MRS broth and incubated at 22°C for 1-3 days. The grown cultures were streaked on MRS agar plates and the generated single colonies were used for preliminary tests, such as Gram detection test and catalase activity to confirm they belong to the LAB group. Subsequently, the isolates were stored at -80°C using 25% w / v glycerol as cryoprotectant.

[0148]

[0115] Example 2: Fermentation of pea protein

[0149]

[0116] Puris Pea 870™ was fermented with Pediococcus pentosaceus C22B 17 and C25R2 strains for 24 hours at 28°C under static conditions. The inoculum size was 1% of fully grown culture at the exponential growth phase.

[0150]

[0117] The sensory analysis of the fermented pea proteins samples are provided in Figure 1, which shows the fermented pea protein suspensions were perceived as much less or moderately less green, beany, pea-like, sour and bitter compared to the control (unfermented pea protein suspension). Additionally, the fermented pea samples were more neutral in terms of taste, slightly sweeter, reminiscent of fresh dough, flour mixture, with improved viscosity, but sightly chalkier mouthfeel.

[0151]

[0118] The chemical analysis of the fermented pea proteins are provided in Figures 2 and 3.

[0119] The “yes” values in the legend of Figure 2 reflects samples inoculated with Pediococcus pentosaceus C25R2. The “no” values in the legend of Figure 2 reflects samples that are not inoculated (blank experiment). The “control” value is a raw Puris Pea 870 protein (control).

[0152]

[0120] Figure 2 shows that replicates of inoculated samples, non-inoculated samples and controls are grouped, except for one outlier for the group 'No, Oh' (in orange). This indicates that method variability, coming from sample preparation and analysis, is low. Samples are separated from controls. The method is then capable of statistically distinguishing samples with known different composition. The first principle component captures about 45% of the total variability and is clearly associated with the chemical modifications that occur for Ferm entate 1, i.e. inoculated after 0 h to the far right, inoculated after 8 h shifted to the left, and inoculated after 24 h to the far left. Replicates from group 'Yes, 24h' are more clearly separated from the rest of the samples. This indicates that at the end of fermentation, the composition has changed in a bigger proportion compared to the first 8 h. The second principal component represents 27% of the variability and seems to differentiate among the blank experiment and control samples that’s not directly relevant to the fermentation process.

[0153]

[0121] The volcano plot shown in Figure 3 indicates the compounds that are down regulated or up regulated in samples after 24 h of fermentation, meaning that the relative amount is significantly lower or higher compared to the 0 h sample, respectively. Each dot corresponds to a single component. Compound concentration in the green box increased in 24h versus Oh, they are produced during fermentation by a significant amount. The increase in concentration could be the result of accumulation of the compound and not necessarily from a higher production rate. It could be attributed to a lower consumption rate in a downstream metabolic pathway. The overall trend would be an increase in the concentration (accumulation) but not because of production but rather due to a constant production and a lower consumption rate. Compound concentration in the red box decreased in 24h versus Oh, they are 'consumed' during fermentation by a significant amount, the decrease in the concentration could be a result of a higher consumption rate or downregulation of the upstream reaction and a constant consumption rate (draining) leading eventually to a lower concentration. In this case there are 193 produced compounds and 147 consumed compounds. Compounds in the 2 colored box and highlighted in blue are selected and reported in the list of compounds.

[0154]

[0122] Among the 340 compounds, families of compounds were identified. Without wishing to be bound by theory, some amino acids are produced during fermentation of the pea protein such as lysine and glutamine. Some amino acids are consumed during fermentation, such as leucine and tyrosine. Several carbohydrates are consumed. Some organic acids are produced, among them are leucic acid and adipic acid.

[0123] To capture the less hydrophilic compounds, samples were extracted with a mixture of acetonitrile and water. Statistical separation between group of replicates for organic / water extract of Fermentate 1 is shown on Figure 4.

[0155]

[0124] Figure 4 shows that replicates of inoculated samples, non-inoculated samples and controls are grouped, but more scattered compared to the water extraction of the LAB fermentation samples. The method variability is slightly higher in this case. Similarly, as in the water extract, samples are separated from controls. Replicates from groups 'Yes, Oh' and 'Yes 8h' are not completely separated. This can indicate that the composition has been less modified between 0 hour and 8 hours for less hydrophilic compounds. This can also be due to the higher variability. Replicates from group 'Yes, 24h' are separated from the groups 'Yes, Oh' and 'Yes 8h'. Replicates from group 'Yes, 24h' are more clearly separated from the rest of the samples. As for water extract, this indicates that at the end of fermentation, the composition has changed in a bigger proportion compared to other time points, even with a higher variability. In figure 3, PC 1 shows the chemical change due to fermentation (45%) while PC 2 only captured differences from controls (27%). In this case controls introduce a lot of variability to the system. An improvement to the protocol could be to prepare a pooled sample (containing equal amounts of each sample) as control. This would center the control in the middle of the PCA graph and group the samples around it.

[0156]

[0125] The volcano plot shown in Figure 5 indicates the compounds that are down regulated or up regulated.

[0157]

[0126] The volcano plot (Figure 5) shows 222 produced compounds and 133 consumed compounds. Similar as for the water extract, several amino acids are found to be either produced or consumed during fermentation. Unlike in the previous case, some carbohydrates are also produced, possibly hexose and ribose or any of their isomers since isomers cannot be distinguished using this method.

[0158] Table 3. Volatiles that decreased significantly after 24 h of fermentation using bacteria cultures*

[0159] *Significance test based on a repeated measures ANOVA with a p-value < 0.05

[0160] Table 4. Volatiles that increased significantly after 24 h of fermentation using bacteria cultures*

[0161] *Significance test based on a repeated measures ANOVA with a p-value < 0.05

[0162]

[0127] Example 3: Production of plant-based alternatives to yoghurt

[0128] A single ingredient mix was made with the below ingredients to produce a white mass which was subsequently fermented with different strain-temperature combinations.

[0163]

[0129] Pea protein (Puris 2.0 or Puris 870) was hydrated with 70% of the total water at 65°C for 5 minutes. The remaining dry ingredients were added to the hydrated pea protein (Puris 2.0 or Puris 870). The remaining 30% of hot water was then added. Melted coconut fat was then added.

[0164]

[0130] The ingredients were mixed under high shear for a minimum of 5 minutes.

[0165]

[0131] The resulting mixture was subjected to a heat treatment: a. Preheating to 70°C before homogenization b. Heat treatment at 95°C for 5 minutes with tubular heating. c. Cooldown to 37°C to reach collecting temperature.

[0166]

[0132] The mixture was collected in fermentation buckets and then fermented as outlined below.

[0167]

[0133] Finally, the yoghurt was smoothed with Silverson high-shear mixer.

[0168]

[0134] Example 4: Sensory data

[0169] QDA was used to determine the effect on sensory characteristics of plant based yogurt of a strain of lactic acid bacterium (Pediococcus pentosaceus C22B17) compared to reference strain (Vega Vibe™ - a commercial starter culture provided by Chr. Hansen A / S which contains a combination of LAB (Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus paracasei subsp. Paracasei and Streptococcus thermophilus') and a mixture of Pediococcus pentosaceus C22B17 and the Vega Vibe™ commercial starter culture at a 50:50 ratio based on two types of Puris pea protein (Puris 2.0 and Puris 870).

[0170] Number of panelists: 9-13 (external trained panel)

[0171] Repetitions: 3

[0172] Serving Procedure:

[0173] ■ Sample presentation: randomized order

[0174] ■ Samples served blind with a 3 -digit code

[0175] ■ Quantity served: ~30ml in plastic cup

[0176] Testing Protocol:

[0177] ■ Samples are swallowed

[0178] ■ Samples are served immediately from the fridge, around 10 °C.

[0179] ■ Break of at least 5 minutes between samples

[0180] ■ 4 products in one session Data collection: EyeQuestion

[0181] Results:

[0182]

[0135] Products associated with the same letters are not significantly different at 10% using Tukey post hoc test. The model considered for the Anova is: Attribute~Product+assessor+replica+Product:assessor+Product:replica. Letters represent groups

[0183]

[0136] The sensory data shown in the table above and in Figures 6-10 confirms that a yoghurt made with pea protein fermented by Pediococcus pentosaceus C22B17 has a clear improvement on the sensory characteristics of the yogurt compared to yoghurts fermented with a commercial starter culture (Vega Vibe).

[0184]

[0137] A plant-based alternative yoghurt (PBAY) yoghurt fermented with Pediococcus pentosaceus C22B17has less overall odor intensity, sour smell, green smell, green taste and sour taste and more old cheese smell and taste, nutty taste, sweet taste and bitter taste compared to a PBAY yoghurt fermented with Vega Vibe™.

[0185]

[0138] In particular, sourness (both taste and aftertaste) is highest for PBAY compositions fermented with Vega Vibe, intermediate for PBAY compositions fermented with the mixture of LAB and lowest for PBAY compositions fermented with Pediococcus pentosaceus C22B17. Sweetness is exactly opposite: lowest for for PBAY compositions fermented with Vega Vibe, intermediate for PBAY compositions fermented with the mixture of LAB and highest for PBAY compositions fermented with Pediococcus pentosaceus C22B17. Texture characteristics mostly show the PBAY compositions fermented with the mixture of LAB having higher values than both PBAY compositions fermented with Pediococcus and PBAY compositions fermented with Vega Vibe. It was found that PBAY compositions fermented with the mixture of LAB has a more creamy and thick mouthfeel and is more cohesive and dense than both PBAY compositions fermented with Pediococcus and PBAY compositions fermented with Vega Vibe. The exception is prickling mouthfeel which is highest for PBAY compositions fermented with Vega Vibe, intermediate for PBAY compositions fermented with the mixture of LAB and lowest for PBAY compositions fermented with Pediococcus.

[0186]

[0139] The features disclosed in the foregoing description, or the following claims, or the accompanying drawings, expressed in their specific forms or in terms of a means for performing the disclosed function, or a method or process for attaining the disclosed result, as appropriate, may, separately, or in any combination of such features, be utilized for realizing the invention in diverse forms thereof.

[0187]

[0140] Although certain example aspects of the invention have been described, the scope of the appended claims is not intended to be limited solely to these examples. The claims are to be construed literally, purposively, and / or to encompass equivalents

Claims

CLAIMS1. A composition comprising Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545 or Pediococcus pentosaceus C25R2 strain obtained or derived BCCM Accession No. LMG P-33546.

2. A starter culture for fermentation of a foodstuff, the starter culture comprising Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545 or Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546.

3. The starter culture of claim 2, wherein the foodstuff is a dairy alternative, optionally, wherein the dairy alternative is a yoghurt alternative.

4. Use of Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545 or Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546 as a starter culture for fermentation of a foodstuff.

5. The use of claim 4, wherein the foodstuff is a dairy alternative, optionally, wherein the dairy alternative is a yoghurt alternative.

6. A fermented plant protein, wherein the plant protein is fermented by lactic acid bacteria, wherein the lactic acid bacteria comprises Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545 or Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546.

7. The fermented plant protein of claim 6, wherein the plant protein is derived from legumes, optionally, wherein the plant protein is a pea protein, optionally, wherein the pea protein is a pea protein isolate; optionally, wherein the pea protein isolate is a hydrolysed pea protein; or, wherein the pea protein is a pea protein concentrate; and / or wherein the lactic acid bacteria further comprises lactic acid bacteria selected from the group consisting of: Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacteriumanimalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus and any combination thereof.

8. A composition comprising: plant protein; and lactic acid bacteria, wherein the lactic acid bacteria comprises Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P-33545 or Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546.

9. The composition of claim 8, wherein the plant protein is derived from legumes; optionally, wherein the plant protein is a pea protein, optionally, wherein the pea protein is a pea protein isolate, optionally, wherein the pea protein isolate is a hydrolysed pea protein; optionally, wherein the pea protein is a pea protein concentrate, and / or wherein the lactic acid bacteria further comprises lactic acid bacteria selected from the group consisting of: Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus and any combination thereof.

10. The composition of claim 8 or 9, wherein the composition further comprises a sugar or sugars, a fat or fats, a liquid or liquids, or a combination thereof, optionally, wherein the liquid is water, syrup, juice or a plant-based or non-dairy alternative to milk, optionally, wherein the liquid is water; and / or, wherein the sugar is selected from the group consisting of sucrose, glucose, dextrose, fructose, maltose, lactose and / or galactose and any combination thereof; and / or, wherein the fat is selected from the group consisting of vegetable fat or vegetable oil; optionally, wherein the vegetable oil is coconut oil, sunflower oil, rapeseed oil, palm oil, shea oil, cocoa oil and any combination thereof; and / or, further comprising one or more emulsifiers, flavourings, thickeners, colourings, additives, probiotics or a combination thereof; and / or, wherein the composition is a fermented composition; and / or, wherein the plant protein is a fermented plant protein; and / or, wherein the composition has one or more of the following characteristics:a. 0.5-12 grams of protein per 100 grams of composition; b. 1-15 grams of carbohydrates per 100 grams of composition; c. 0-10 grams of fat per 100 grams of composition; and / or d. 0-150 mg of calcium per 100 grams of composition; and / or, wherein the composition is a foodstuff; optionally, wherein the foodstuff is a yoghurt alternative.

11. A process for preparing the fermented plant protein of claims 6 or 7 or the composition of claims 8-10, the process comprising: a) Providing an aqueous composition comprising a plant protein; b) Providing lactic acid bacteria, wherein the lactic acid bacteria comprises Pediococcus pentosaceus C22B17 strain obtained or derived from BCCM Accession No. LMG P- 33545, or, Pediococcus pentosaceus C25R2 strain obtained or derived from BCCM Accession No. LMG P-33546.

12. The process of claim 11 for preparing a fermented plant protein, wherein step (b) comprises fermenting the composition with the lactic acid bacteria to provide a fermented plant protein; and / or, wherein the plant protein is derived from legumes; optionally, wherein the plant protein is a pea protein, optionally, wherein the pea protein is a pea protein isolate, optionally, wherein the pea protein isolate is a hydrolysed pea protein; optionally, wherein the pea protein is a pea protein concentrate; and / or, wherein step (b) is carried out at 25-45°C, optionally, wherein step (b) is carried out at 28°C; and / or, wherein the lactic acid bacteria further comprises lactic acid bacteria selected from the group consisting of: Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus and any combination thereof, optionally, wherein Pediococcus pentosaceus and Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, or Lactobacillus helveticus and any combination thereof are provided in a 50:50 ratio; optionally, wherein step (b) is carried out at 25-45°C, optionally, wherein step (b) is carried out at 28-37°C.

3. The process of claim 11 for preparing a composition, wherein the plant protein is derived from legumes, optionally, wherein the plant protein is a pea protein; optionally, wherein the pea protein is a pea protein isolate; optionally, wherein the pea protein isolate is a hydrolysed pea protein; or, wherein the pea protein is a pea protein concentrate; and / or, wherein step (b) is carried out at 25-45°C; optionally, wherein step (b) is carried out at 28°C; and / or, wherein the lactic acid bacteria in step (b) ferments the aqueous composition comprising a plant protein; and / or, further comprising providing lactic acid bacteria selected from the group consisting of: Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus and any combination thereof; optionally wherein Pediococcus pentosaceus and Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. Bulgaricus, Streptococcus thermophilus, Bifidobacterium species, Biofidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus casei, Lactobacillus plantarum, or Lactobacillus helveticus and any combination thereof are provided in a 50:50 ratio; optionally, wherein step (b) is carried out at 25-45°C; optionally, wherein step (b) is carried out at 28-37°C; and / or, wherein the plant protein is hydrated; and / or, wherein the composition is subjected to mixing after step (a) but before step (b); and / or, wherein the composition is homogenized after step (a) but before step (b); and / or, wherein the composition is subjected to a heat treatment after step (a) but before step (b) to pasteurise the composition; optionally, wherein the heat treatment is 2-10 minutes at a temperature of 80-100°C; and / or wherein the composition is cooled before step (b); optionally, wherein the composition is cooled to the fermentation temperature of the lactic acid bacteria; optionally, wherein the composition is cooled to 25-45°C; and / or, wherein the process further comprises (c) smoothening the composition; and / or, wherein step (a) further comprises adding one or more additional ingredients to the aqueous composition comprising a plant protein; and / or, wherein the one or more additional ingredients include a sugar or sugars, a fat or fats, a liquid or liquids, emulsifiers, flavourings, thickeners, colourings, additives, probiotics or a combination thereof, optionally, wherein the liquid is water, syrup, juice or a plant-based or non-dairy alternative to milk, optionally, wherein the liquid is water; and / or, wherein the sugar is selected from the group consisting of sucrose, glucose, dextrose, fructose, maltose, lactose and / or galactose and anycombination thereof; and / or, wherein the fat is selected from the group consisting of vegetable fat or vegetable oil, optionally, wherein the vegetable oil is coconut oil, sunflower oil, rapeseed oil, palm oil, shea oil, cocoa oil and any combination thereof.

14. A food product comprising the fermented plant protein of claims 6 or 7 or the composition of any one of claims 8-10.

15. The food product of claim 14, wherein the product is a dairy alternative, optionally, wherein the dairy alternative is a yoghurt alternative.

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